PE Anti-Human CD45 Antibody[HI30]
Price: $ 63
Price: $ 22
Price: $ 98
- Application: FCM
- Isotype: Mouse IgG1, κ
- Host: Mouse
For research use only. Order now, ship in 3 days
Background | CD45 is a 180 - 240 kD single chain type I membrane glycoprotein also known as leukocyte common antigen (LCA) and T200. It is a tyrosine phosphatase expressed on the plasma membrane of all hematopoietic cells, except erythrocytes or platelets. CD45 is a signaling molecule that regulates a variety of cellular processes including cell growth, differentiation, cell cycle, and oncogenic transformation. CD45 plays a critical role in T and B cell antigen receptor-mediated activation by dephosphorylating substrates including p56Lck, p59Fyn, and other Src family kinases. CD45 non-covalently associates with lymphocyte phosphatase-associated phosphoprotein (LPAP) on T and B lymphocytes. CD45 has been reported to bind galectin-1 and to be associated with several other cell surface antigens including CD1, CD2, CD3, and CD4. |
Alternate Names | Receptor-type tyrosine-protein phosphatase C;Ptprc;L-CA;Ly-5;T200;CD45; |
Swissprot | |
Clone No | |
Application |
FCM
|
Host | Mouse |
Reactivity | Human |
Isotype | Mouse IgG1, κ |
Isotype Control | |
Form | Liquid |
Conjugation | PE |
Conjugation information | PE is designed to be excited by the Blue (488 nm), Green (532 nm) and Yellow-Green (561 nm) lasers and detected using an optical filter centered near 575 nm (e.g., a 585/42 nm bandpass filter). |
Spectrum | |
Storage Buffer | Phosphate buffered solution, pH 7.2, containing 0.09% stabilizer and 1% protein protectant. |
Storage | This product can be stored at 2-8°C for 12 months. Please protected from prolonged exposure to light and do not freeze. |
Expiration date | 12 months |
Shipping | Ice bag |
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Q1:Is it avaliable to store FCM antibodies at -20°C and thaw before use?
The stability of the flow antibody can be different from each other based on differnet fluorescein. It is recommended to verify the freeze-thawed antibody through pre-experiments before determining. However, it is better to avoid the freeze-thawed cycling in any FCM antibody.
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Q2:Is it necessary to take blocking? Or for some specific samples?
Blocking process is required when detecting macrophages, dendritic cells, NK cells. Fc receptors can be expressed on macrophages, dendritic cells, NK cells, etc. In the process of antibody staining in flow assay, Fc segment of FCM antibody will bind to Fc receptors on cell surface, end up with non-specific staining and lead to false positives signals. Antibodies can be incubated directly after blocking without washing.
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Q3:Why centrifuge before use?
During the transportation of antibodies, antibodies will stick to the tube wall or cap due to turbulence. So after receiving the antibodies, moderate centrifugation will collect the antibodies on the tube wall or cap to the bottom of the tube to avoid the loss of antibodies.
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Q4:What auxiliary reagents are needed for staining?
For samples with erythrocyte, ACK lysis buffer (E-CK-A105) is needed; For cells that are rich in Fc receptors, such as macrophages, Fc receptors blocking is necessary before staining with flow Antibody to reduce the non-specific signal. At present, we can provide human and mouse blocking agent, E-AB-F1236A Purified Anti-Human CD16 Antibody[3G8]. E-AB-F0997A Purified Anti-Mouse CD16/32 Antibody[2.4G2]. Cell staining buffer (E-CK-A107) is required in the process of cell staining. For the detection of intracellular indexes, a fixation & permeabilization kit is needed (E-CK-A109) ; and for the detection of intranuclear indexes, a specific staining kit (E-CK-A108) is required. Dead cell dyes are also used for flow cytometry of tissue samples such as tumors.
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Q5:How should experimental groups/controls be set?
Blank control: used to set the voltage of each channel. Isotype control: Isotype control antibodies are used as the basis for determining negative and positive cells. It is necessart as a gating helper especially for the indicators with low expression or continuous expression. Single color control: In a multicolor experiment, single color control is needed to adjust the fluorescence compensation if there is interference between the different channels. FMO control: FMO control, also known as fluorescence reduction control, refers to the multi-color experiment. FMO control is applied to observe the comprehensive effect of all related fluoresents to the target channel by removing the correspinding signal.
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Q6:If there is only 1×10^5 cells instead of 1×10^6, can the antibody dosage be reduced?
The amount of antibody is related to the incubation system, if the cell suspension volume is still 100 μL, the amount of antibody remains the same. Researchers can reduce the amount to save antibody in low cell number condition by reducing the cell suspension volume. It is recommended to use the recommended number of cells for the experiment, if the number of cells is too large, it will lead to insufficient antibody dosage, resulting in false negative; if the number of cells is too small, especially when detecting intracellular or intranuclear indicators, a large number of centrifugation operations will lose a lot of cells, resulting in insufficient number of cells for final detection.
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Q7:How to dilute the Test package antibodies?
The usage of test-package antibodies is well designed and verified, there is no need for an extra dilution. The usage amout of 1 test is 5 μL of antibodies per 100μL cell suspension (containing 1x10^6 cells).
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Q8:What is the difference between the test-package and the weight-package of flow antibody products?
The usage of test-package antibodies is well designed and verified there is no need for an extra dilution before use. The weight-package antibody has a higher concentration, and requires a titration process for a suitable usage amount.
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Q9:What does Isotype Control do? How to choose a suitable isotype control antibody?
Isotype control antibodies are used as the basis for determining negative and positive cells. It is necessart as a gating helper especially for the indicators with low expression or continuous expression. The Isotype control was purified from the serum of non-immunized animals, it should be the same species source, same immunoglobulin and subtype, same fluorescein label, same dose and concentration as the stained monoclonal antibody.